Calf thymus high mobility group proteins are nonenzymatically glycated but not significantly glycosylated.
Over the past decade, there have been many reports suggesting the presence of complex carbohydrates on nuclear and cytoplasmic proteins in mammalian cells. Some of the most often cited of these reports deal with the glycosylation of the high mobility group (HMG) proteins. These are relatively abundant chromosomal proteins that are known to be associated with nucleosomes and actively transcribed regions of chromatin. The original report describing HMG protein glycosylation presented several lines of evidence suggesting that these proteins are glycosylated, including carbohydrate compositional analysis and periodic-acid Schiff staining. We have attempted to repeat these observations with more highly purified protein than was utilized in the original study. Using carbohydrate compositional analysis performed by high pH anion exchange chromatography coupled to pulsed-amperometric detection, we saw no evidence for significant glycosylation of these proteins. In addition, we found no evidence for the presence of O-GlcNAc, a well known form of nuclear glycosylation. The HMG proteins did react with periodate, suggesting the presence of a modification containing cis-diols on the protein. Several tryptic peptides isolated from HMG 14 and 17 which retained the periodate reactivity had in common lysine residues, suggesting a potential modification of the straightepsilon-amino groups of lysines such as nonenzymatic glycation. Western blot analysis of the HMG proteins using anti-advanced glycation endproducts (AGE) antibodies confirmed the presence of glycation products on the HMG proteins.
Medina
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Department of Biochemistry and Cell Biology, Institute for Cell and Developmental Biology, State University of New York at Stony Brook 11794-5215, USA.
Haltiwanger
R S
RS
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D005947:Glucose / Q000378:metabolism*; D006031:Glycosylation; D006609:High Mobility Group Proteins / Q000302:isolation & purification / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D010503:Periodic Acid-Schiff Reaction; D013950:Thymus Gland / Q000378:metabolism*
D002241:Carbohydrates; D006609:High Mobility Group Proteins; D010446:Peptide Fragments; D005947:Glucose; D000117:Acetylglucosamine